CRISPR/Cas9-mediated knockout of Lcn2 effectively enhanced CDDP-induced apoptosis and reduced cell migration capacity of PC3 cells

Rahimi, Sina; Roushandeh, Amaneh Mohammadi; Ebrahimi, Ammar; Samadani, Ali Akbar; Kuwahara, Yoshikazu; Roudkenar, Mehryar Habibi

doi:10.1016/j.lfs.2019.116586

Cited by 37 publications

(28 citation statements)

References 31 publications

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“…4 B). LCN2 knockout by CRISPER/Cas9 plasmid decreased cell proliferation and increased sensitivity to cisplatin in prostate cancer cells 67 . LCN2-deficent mice showed increased intracellular labile iron, and were highly sensitive to LPS-induced mortality, associated with increased cell apoptosis and upregulation of proinflammatory gene expression 68 .…”

Section: Discussionmentioning

confidence: 96%

Subtilase cytotoxin induces a novel form of Lipocalin 2, which promotes Shiga-toxigenic Escherichia coli survival

Yahiro¹,

Ogura²,

Goto³

et al. 2020

Sci Rep

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Shiga-toxigenic Escherichia coli (STEC) infection causes severe bloody diarrhea, renal failure, and hemolytic uremic syndrome. Recent studies showed global increases in Locus for Enterocyte Effacement (LEE)-negative STEC infection. Some LEE-negative STEC produce Subtilase cytotoxin (SubAB), which cleaves endoplasmic reticulum (ER) chaperone protein BiP, inducing ER stress and apoptotic cell death. In this study, we report that SubAB induces expression of a novel form of Lipocalin-2 (LCN2), and describe its biological activity and effects on apoptotic cell death. SubAB induced expression of a novel LCN2, which was regulated by PRKR-like endoplasmic reticulum kinase via the C/EBP homologous protein pathway. SubAB-induced novel-sized LCN2 was not secreted into the culture supernatant. Increased intracellular iron level by addition of holo-transferrin or FeCl3 suppressed SubAB-induced PARP cleavage. Normal-sized FLAG-tagged LCN2 suppressed STEC growth, but this effect was not seen in the presence of SubAB- or tunicamycin-induced unglycosylated FLAG-tagged LCN2. Our study demonstrates that SubAB-induced novel-sized LCN2 does not have anti-STEC activity, suggesting that SubAB plays a crucial role in the survival of LEE-negative STEC as well as inducing apoptosis of the host cells.

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Section: Discussionmentioning

confidence: 96%

Subtilase cytotoxin induces a novel form of Lipocalin 2, which promotes Shiga-toxigenic Escherichia coli survival

Yahiro¹,

Ogura²,

Goto³

et al. 2020

Sci Rep

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“…Lipocalin‐2/NGAL has been shown to be overexpressed in many solid cancers. In particular, for PCa, its overexpression increased migration and invasion 31,32 ; whereas attenuation of its expression in the PCa cells decreased these processes 33,34 . Lipocalin‐2 was even considered as a screening test for PCa 35 .…”

Section: Discussionmentioning

confidence: 99%

“…whereas attenuation of its expression in the PCa cells decreased these processes. 33,34 Lipocalin-2 was even considered as a screening test for PCa. 35 Microseminoprotein-β (MSMB)/Prostate secretory protein of 94 amino acids (PSP94), primarily found in the prostatic secretion, was originally isolated and purified from human seminal plasma.…”

Section: Validation Of Differential Expression Of Selected Genes Anmentioning

confidence: 99%

Proteomic and transcriptomic profiling of Pten gene‐knockout mouse model of prostate cancer

Zhang

Kim

et al. 2020

The Prostate

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Background The prostate‐specific phosphatase and tensin homolog deleted on chromosome 10 (Pten) gene‐conditional knockout (KO) mouse carcinogenesis model is highly desirable for studies of prostate cancer biology and chemoprevention due to its close resemblance of primary molecular defect and many histopathological features of human prostate cancer including androgen response and disease progression from prostatic intraepithelial neoplasia to invasive adenocarcinoma. Here, we profiled the proteome and transcriptome of the Pten‐KO mouse prostate tumors for global macromolecular expression alterations for signaling changes and biomarker signatures. Methods For proteomics, four pairs of whole prostates from tissue‐specific conditional knockout Pten‐KO mice (12‐15 weeks of age) and their respective wild‐type littermates housed in the same cages were analyzed by 8‐plex isobaric tags for relative and absolute quantitation iTRAQ. For microarray transcriptomic analysis, three additional matched pairs of prostate/tumor specimens from respective mice at 20 to 22 weeks of age were used. Real‐time quantitative reverse transcription‐polymerase chain reaction was used to verify the trends of protein and RNA expression changes. Gene Set Enrichment Analysis and Ingenuity Pathway Analysis were carried out for bioinformatic characterizations of pathways and networks. Results At the macromolecular level, proteomic and transcriptomic analyses complement and cross‐validate to reveal overexpression signatures including inflammation and immune alterations, in particular, neutrophil/myeloid lineage suppressor cell features, chromatin/histones, ion and nutrient transporters, and select glutathione peroxidases and transferases in Pten‐KO prostate tumors. Suppressed expression patterns in the Pten‐KO prostate tumors included glandular differentiation such as secretory proteins and androgen receptor targets, smooth muscle features, and endoplasmic reticulum stress proteins. Bioinformatic analyses identified immune and inflammation responses as the most profound macromolecular landscape changes, and the predicted key nodal activities through Akt, nuclear factor‐kappaB, and P53 in the Pten‐KO prostate tumor. Comparison with other genetically modified mouse prostate carcinogenesis models revealed notable molecular distinctions, especially the dominance of immune and inflammation features in the Pten‐KO prostate tumors. Conclusions Our work identified prominent macromolecular signatures and key nodal molecules that help to illuminate the patho‐ and immunobiology of Pten‐loss driven prostate cancer and can facilitate the choice of biomarkers for chemoprevention and interception studies in this clinically relevant mouse prostate cancer model.

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“…We transfected the CRISPR-Cas9 plasmids carrying PUM1 or DDX5 to SW480R or Caco-2R cell lines with Lipofectamine 2000 (11668019, Thermo Fisher Scientific). The stably transfected cells were selected in the presence of 5 μg/ml puromycin for 2 weeks ( Rahimi et al, 2019 ). DDX5 overexpression was also performed with pSIN-GFP carrying DDX5 coding sequence (CDS) without 3′ untranslated region (UTR) and transfected to corresponding colon cancer cell lines by Lipofectamine 2000 according to the manufacturer’s instruction.…”

Section: Methodsmentioning

confidence: 99%

PUM1 Is Overexpressed in Colon Cancer Cells With Acquired Resistance to Cetuximab

Liu

Cheng

Chen

et al. 2021

Front. Cell Dev. Biol.

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BackgroundCetuximab is an effective antibody to treat colorectal cancer (CRC) by targeting the epidermal growth factor receptor (EGFR). However, the mechanisms of acquired resistance to cetuximab therapy, especially in patients without identifiable gene mutations, are not fully understood.MethodsOur study investigated the role of pumilio RNA-binding family member 1 (PUM1) in cetuximab resistance. We established cetuximab-resistant colon cancer cell lines SW480R and Caco-2R and knocked out PUM1 and DEAD-box helicase 5 (DDX5) with the clustered regularly interspaced short palindromic repeats (CRISPR)-caspase 9 (Cas9) system. To check cell proliferation, we used Cell Counting Kit-8. We performed qPCR and immunoblot to examine the levels of mRNAs and proteins for each cell line.ResultsOur data showed that PUM1 was upregulated in SW480R and Caco-2R cells with increased protein levels and cell proliferation, and PUM1 knockout reduced cell viability in the presence of cetuximab. We also found that PUM1 interacted with DDX5 in 3′ untranslated region (UTR) and positively regulated its mRNA expression. Furthermore, suppression of DDX5 also decreased the proliferation of SW480R and Caco-2R cells.ConclusionOur study suggests that PUM1 positively regulates DDX5 and acts as a promoter in cetuximab-resistant colon cancer cells.

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CRISPR/Cas9-mediated knockout of Lcn2 effectively enhanced CDDP-induced apoptosis and reduced cell migration capacity of PC3 cells

Cited by 37 publications

References 31 publications

Subtilase cytotoxin induces a novel form of Lipocalin 2, which promotes Shiga-toxigenic Escherichia coli survival

Subtilase cytotoxin induces a novel form of Lipocalin 2, which promotes Shiga-toxigenic Escherichia coli survival

Proteomic and transcriptomic profiling of Pten gene‐knockout mouse model of prostate cancer

PUM1 Is Overexpressed in Colon Cancer Cells With Acquired Resistance to Cetuximab

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